Kit for extracting aquatic animal fecal like bacteria DNA and method thereof
A technology for aquatic animals and a kit, which is applied in the field of extracting bacterial DNA from feces samples, can solve the problems of extracting DNA with many impurities and incomplete extraction, and achieve the effect of comprehensive information, rapid extraction method and high yield
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Embodiment 1
[0034] Make the bacterial DNA extraction kit of aquatic animal feces samples according to the following formula:
[0035] Reagent A: Contains 70% (V / V) ethanol and 0.5% (V / V) Triton X-100.
[0036] Reagent B: Contains 50 mM Tris-Cl (pH 8.0), 50 mM EDTA, 50 mM NaCl, 2% (W / V) of PVP, 1% (W / V) of CTAB and 3% (W / V) of SDS.
[0037] Reagent C: RNAase concentration is 20mg / ml.
[0038] Reagent D: Contains 6.5M GuSCN (guanidine isothiocyanate) and 100mM Tris, adjust pH to 6.
[0039] Reagent E: prepared by mixing 10 mM Tris-Cl (pH 7.5) and absolute ethanol at a ratio of 1:4.
[0040] Reagent F: 10 mM Tris-Cl (pH 8.0).
[0041] DNA adsorption column: silica gel column
[0042] Follow the procedure below to extract bacterial DNA from aquatic animal fecal samples:
[0043] 1. Sample pretreatment
[0044] Take 200 mg of fish feces sample, add 400 μl of reagent A, place on ice for 30 min, centrifuge at 12000 g for 5 min.
[0045] 2. Bacterial Lysis
[0046] Discard the supernatant, ...
Embodiment 2
[0054] Various reagents were prepared according to the following formula:
[0055] Reagent A: Contains 75% (V / V) ethanol and 0.1% (V / V) Triton X-100.
[0056] Reagent B: Contains 200 mM Tris-Cl (pH 8.0), 100 mM EDTA, 100 mM NaCl, 1% (W / V) of PVP, 0.5% (W / V) of CTAB and 2% (W / V) of SDS.
[0057] Reagent C: RNAase concentration is 10 mg / ml.
[0058] Reagent D: Contains 6.5M GuSCN (guanidine isothiocyanate) and 50mM Tris, adjust pH to 6.
[0059] Reagent E: prepared by mixing 10mM Tris-Cl (pH 7.5) and absolute ethanol 1:4.
[0060] Reagent F: ddH 2 O.
[0061] DNA adsorption column: silica gel column
[0062] Follow the procedure below to extract bacterial DNA from aquatic animal fecal samples:
[0063] 1. Sample pretreatment
[0064] Take 100mg of prawn feces, add 200μl reagent A, place on ice for 20min, centrifuge at 12000g for 5min.
[0065] 2. Bacterial Lysis
[0066] Discard the supernatant, add 200 μl of reagent B and 4 μl of reagent C to the pellet, stir well with...
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